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Manganese-based nano-enzyme as ferroptosis inhibitor and application of manganese-based nano-enzyme in liver injury

A nano-enzyme and ferroptosis technology, which is applied in the field of medical applications, can solve the problems of high price, ineffective effect, and difficulty in controlling the dosage, and achieve the effect of alleviating drug-induced liver injury, inhibiting ferroptosis, and stabilizing the inhibitory effect for a long time

Active Publication Date: 2022-05-27
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But the price is relatively expensive, and the effect is not obvious when inhibiting ferroptosis induced by certain drugs; the effect of iron chelators defetoxamine (DFO) and defriprone (DFP) is not particularly significant, and the dosage is difficult take control

Method used

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  • Manganese-based nano-enzyme as ferroptosis inhibitor and application of manganese-based nano-enzyme in liver injury
  • Manganese-based nano-enzyme as ferroptosis inhibitor and application of manganese-based nano-enzyme in liver injury
  • Manganese-based nano-enzyme as ferroptosis inhibitor and application of manganese-based nano-enzyme in liver injury

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation method of manganese-based nanozyme preparation is as follows:

[0027] Step S1: the MnSO 4 ·H 2 Dissolve 0.425 g of O in 2.5 mL of pure water, and dissolve 4.5 g of polyacrylic acid (weight-average molecular weight of 1800) in 5 mL of pure water; after mixing the above manganese sulfate solution and polyacrylic acid solution, put it on a vortexer at 2500 rpm Mix for 15 min to obtain system A solution;

[0028] Step S2: The mixed solution was added dropwise to 15 mL of 30 wt % ammonia water, and stirred at 500 rpm on a magnetic stirrer for 24 h; the stirred solution was put into a 50 mL polytetrafluoroethylene reaction kettle at 120° C. The hydrothermal reaction was carried out for 24 h; the reacted solution was divided into 2 mL centrifuge tubes, centrifuged at 4000 × g for 1 h at room temperature, the supernatant was taken, dialyzed with a dialysis bag (MW 3500) for 24 h, and the water was changed every 8 h to obtain Aqueous formulations containing m...

Embodiment 2

[0030] Screening of suitable concentration of manganese-based nanozyme preparations (Mn-NPs) to inhibit RSL3-induced ferroptosis

[0031] Take logarithmic growth phase MEF cells, trypsin digestion, press 2.5 × 10 per well 5 (500 μl per well) were inoculated into a 24-well plate, cultured for 24 h, the medium was discarded, and replaced with the following medicated medium:

[0032] RSL3 group: RSL3 (ferroptosis inducer) was diluted to 1 μM with cell culture medium;

[0033] Fer group: Ferrostatin was diluted to 10 μM with cell culture medium;

[0034] RSL3+Mn-NPs group: Manganese-based nanozyme preparations were diluted with cell culture medium to 11.2, 22.4, 33.6, 44.8 and 56 μg / ml, and 1 μM RSL3 was simultaneously added to each group on this basis.

[0035] The cell culture medium was DMEM containing 10% FBS and 1% double antibody. After 4 h of treatment, the drug-containing medium was discarded, and 100 μl of 4 μM PI staining solution (diluted in PBS) was added to each we...

Embodiment 3

[0038] Manganese-based nanozyme preparations (Mn-NPs) inhibit RSL3 and Erastin-induced ferroptosis

[0039] Take logarithmic growth phase MEF cells, trypsin digestion, press 2.5 × 10 per well 5 (500 μl per well) were inoculated into a 24-well plate, cultured for 24 h, the medium was discarded, and replaced with the following medicated medium:

[0040] RSL3 group: RSL3 (ferroptosis inducer) was diluted to 1 μM with cell culture medium;

[0041]Erastin group: Erastin (ferroptosis inducer) was diluted to 10 μM with cell culture medium;

[0042] Fer group: Fer was diluted to 10 μM with cell culture medium;

[0043] RSL3+Mn-NPs group: The manganese-based nanozyme preparation was diluted to 56 μg / ml with cell culture medium, and 1 μMRSL3 was added at the same time;

[0044] Erastin+Mn-NPs group: The manganese-based nanozyme preparation was diluted to 56 μg / ml with cell culture medium, and 10 μM Erastin was added at the same time;

[0045] RSL3+Fer group: cell culture medium cont...

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Abstract

The invention relates to the technical field of medical application, and discloses a manganese-based nano-enzyme preparation (Mn-NPs) as a ferroptosis inhibitor and application of the manganese-based nano-enzyme preparation in liver injury. The manganese-based nano-enzyme preparation is applied to mouse embryo fibroblasts and living mice, and it is found that the manganese-based nano-enzyme preparation as a ferroptosis inhibitor can significantly inhibit cell ferroptosis induced by RSL3 and Erastin, has a long-term stable inhibition effect, and also has a relieving effect on acetaminophen (APAP)-induced mouse drug-induced liver injury.

Description

technical field [0001] The invention relates to the technical field of medical applications, in particular to a manganese-based nanozyme as a ferroptosis inhibitor and its application in liver injury. Background technique [0002] Ferroptosis is a new type of programmed cell death that is iron-dependent and different from apoptosis, necrosis and autophagy. It does not have the morphological characteristics of apoptosis, and does not have the phenomena that occur in traditional apoptosis, such as cell shrinkage, chromatin condensation, the formation of apoptotic bodies, and the disintegration of the cytoskeleton. Significant shrinkage of mitochondrial morphology and increased membrane density can be observed, which is absent in apoptosis. The main mechanism of ferroptosis is that under the action of ferrous iron or ester oxygenase, it catalyzes lipid peroxidation of highly expressed unsaturated fatty acids on the cell membrane, thereby inducing cell death; in addition, it is...

Claims

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Application Information

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IPC IPC(8): A61K33/32A61K47/54A61K47/58A61K47/59A61P1/16B22F9/16B82Y5/00
CPCA61K47/58A61K47/59A61K47/542A61K33/32A61P1/16B22F9/16B82Y5/00Y02A50/30
Inventor 吴浩吴洪洪李佳欢单心怡刘家浩
Owner HUAZHONG AGRI UNIV
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